Immunohistochemistry, Immunocytochemistry, Immunofluoresence, Western Blot
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Description
The C-FOS Antibody was raised to synthetic peptide sequence corresponding to human c-fos (4-17) coupled to bovine thyroglobulin with glutaraldehyde. For induction of c-fos protein activity rats were injected with 1.0 ml of 1.5 M NaCl per 100 grams of body weight. Negative control rats were injected with the same volume of normal saline. The ImmunoStar c-fos antiserum was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat paraventricular nucleus and supraoptic nucleus using indirect immunofluorescent and biotin/avidin-HRP techniques. No labeling was seen in negative control rats. Recommended primary dilutions for these methods are 1/200-1/400 in PBS/0.3% Triton X-100 - FITC Technique and 1/4000-1/6000 in PBS/0.3% Triton X-100 - biotin/avidin-HRP Technique. Specificity of the antiserum was demonstrated by blockage of staining in experimental rats by omission of c-fos antibody or by substitution of antibody pre-incubated with synthetic peptide or the conjugate. Immunoblot analysis of mediobasal hypothalamus showed a single band of approximately 55-60 kD.